三七姜挥发油诱导(CNE-2)细胞周期G_1期阻滞及相关基因表达的影响  

Induction of G_1 arrest of CNE-2 cells by volatile oils fromStahlianthus involucratus and effects on related gene expression

作　　者：王蔓莉[1] 邓立东[1] 徐勤[1] 

机构地区：[1]桂林医学院,广西桂林541004

出　　处：《中国医院药学杂志》2015年第5期409-413,共5页Chinese Journal of Hospital Pharmacy

基　　金：广西科技厅重大专项计划资助(编号:桂科重12118005-2-3);广西教育厅重点资助项目(编号:201202ZD067)

摘　　要：目的:研究三七姜挥发油对人鼻咽癌(CNE-2)细胞周期的调控作用,并探讨其可能的机制。方法:体外培养人鼻咽癌细胞株(CNE-2),采用MTT法观察三七姜挥发油对CNE-2细胞增殖的抑制作用;流式细胞术分析三七姜挥发油处理CNE-2细胞后对细胞周期分布的影响;实时荧光定量PCR检测细胞周期调控相关基因的表达水平。结果:不同浓度的三七姜挥发油作用CNE-2细胞24,48,72,96 h后,均能明显抑制细胞增殖(P<0.05),且呈浓度与时间依赖性,其IC50分别为71.12,60.034,53.167,34.75μg·ml-1;三七姜挥发油诱导CNE-2细胞发生G1期阻滞,与阴性对照相组比,200μg·ml-1加药处理CNE-2细胞72 h后G1期细胞增加了13.94%(P<0.01);伴随ATM,CCND1/E1,CDK6,CDC25A,E2F1等mRNA表达水平下调,而Tp53基因表达显著增强(P<0.01)。结论:三七姜挥发油诱导人鼻咽癌CNE-2细胞G1期阻滞、抑制细胞增殖,其作用可能与抑制了介导DNA损伤修复的ATM-Chk2-Cdc25A通路,以及激活p53通路有关,从而抑制CDC25A,cyclinD1/E1、CDK6、E2F1等基因的表达而实现的。OBJECTIVE To investigate the regulating effects of volatile oils fromStahlianthus involucratus on cell cycle of human nasopharyngeal carcinoma cell line CNE-2 in vitro and the underlying potential mechanism.METHODS CNE-2 cells were cultured invitro,and 3-（4,5-dimethylthiazole-2-yl）-2,5-diphenyltetrazolium bromide（MTT）assay was used to observe the inhibiting effects of volatile oil fromStahlianthus involucratus on proliferation of CNE-2 cell lines.The distribution of cell cycle of CNE-2 cells were determined by flow cytometry（FCM）.Expression of cell cycle regulation-related genes were assessed by real-time polymerase chain reaction（RT-PCR）method.RESULTS Incubation with volatile oils fromStahlianthus involucratuson for 24,48,72 and 96 h resulted in a significant inhibition of CNE-2 cell proliferation in a dose and time dependent manner（P〈0.05）,and IC50 values for them were 71.12,60.034,53.167μg·ml^-1 and 34.75μg·ml-1,respectively.Volatile oils fromStahlianthus involucratusoninduced G1 phase arrest,compared with the untreated control group,G1 phase of the cells treated by 200μg·ml^-1 drug increased by 13.94%after 72 h（70.03±1.15）%vs.（56.09±2.12）%（P〈0.01）.The mRNA levels of ATM,CCND1/E1,CDK6,CDC25 A,and E2F1 decreased,while Tp53 elevated（P〈0.01）.CONCLUSIONVolatile oils can potently disturb proliferation of CNE-2 cells in vitro and induce cell cycle arrest at G1 phase,which may be realized by inhibiting ATM-Chk2-Cdc25 A pathway which mediates DNA injury restoration,activating p53 pathway,and then inhibiting expression of genes of CDC25 A,cyclinD1/E1,CDK6 and E2F1.

关 键 词：三七姜挥发油 人鼻咽癌细胞CNE-2 细胞周期阻滞 

分 类 号：R96[医药卫生—药理学]